Lyme borreliosis in humans is a multisystemic disorder caused by infection with the tick-borne spirochete, Borrelia burgdorferi, (Burgdorfer et al. 1982. Science 216:1317-1319; Johnson et al. 1984. Int. J. Syst. Bacteriol. 34:496-497; and Steere et al. 1983. N. Engl. J. Med. 308:733-740). Since the first epidemiological investigations of this disease in south-central Connecticut (Steere et al. 1977. Ann. Intern. Med. 86:685-698 and Steere et al. 1977. Arthritis. Rheum. 20:7-17), human cases of Lyme borreliosis have now been acquired in 43 states of the United States (Centers for Disease Control 1989, Lyme Disease--United States, 1987 and 1988. MMWR 38:668-672), five provinces of Canada, (Centers for Disease Control 1989, Lyme disease--Canada. MMWR 38:677-678), numerous countries throughout Europe and Asia (Ai et al. 1988. Ann. N.Y. Acad. Sci. 539:302-313; Dekonenko et al. 1988. J. Infect. Dis. 158:748-753; and Schmid. 1985. Rev. Infect. Dis. 7:41- 50), and possibly restricted foci in Australia (Stewart et al. 1982. Med. J. Australia 1:139) and Africa (Haberberger et al. 1989. Trans. R. Soc. Trop. Med. Hyg. 83:556 and Stanek et al. 1986. Zentralbl. Bakteriol. Mikrobio. Hyg. [A] 263:491-495). Between 1982-1988, reports of 13,825 cases of Lyme borreliosis were received by the Centers for Disease Control from all 50 states of the United States, (Centers for Disease Control 1989, Lyme Disease--United States, 1987 and 1988. MMWR 38:668-672), making this disease the most prevalent arthropod-borne infection in the country.
With the dramatic increase in awareness, prevalence, and geographical distribution of Lyme borreliosis, a tremendous new demand has been placed on clinical laboratories for serological confirmation of cases, (Magnarelli. 1989. J. Am. Med. Assoc. 262:3464-3465 and Schwartz et al. 1989. J. Am. Med. Assoc. 262:3431-3434) or to rule out this disease in differential diagnoses. However, many potential problems exist with the currently available serological tests for Lyme borreliosis, which may result in either false positive or false negative results (Magnarelli 1989. J. Am. Med. Assoc. 262:3464-3465). Some studies have focused on using flagellar protein of B. burgdorferi to increase the sensitivity of serological tests (Hansen et al. 1989. J. Clin. Microbiol 27:545-551 and Hansen et al. 1988. J. Clin. Microbiol 26:338-346) because earlier studies demonstrated that it appeared to be the 41 kilodalton (kDa) flagellar subunit (flagellin) of the spirochete that generated the earliest antibody response in infected humans (Barbour et al. 1983. J. Clin. Invest. 72:504-515; Coleman et al. 1987. J. Infect. Dis. 155:756-765; and Grodzicki et al. 1988. J. Infect. Dis. 157:790-797). One of two potential problems with using flagellar protein, however, is that flagella of other Borrelia species share epitopes common to the flagella of B. burgdorferi (Barbour et al. 1986. Infect. Immun. 52:549-544). Secondly, in most studies that have screened human sera by immunoblot analysis (Barbour. 1984. Yale J. Biol. Med. 57:581-586; Barbour et al. 1983. J. Clin. Invest. 72:504-515; Coleman et al. 1987. J. Infect. Dis. 155:756-765; Craft et al. 1986. J. Clin. Invest. 78:934-939; and Nadal et al. 1989. Pediatr. Res. 26:377-382), antibodies binding the protein with an apparent migration of 41 kDa have been assumed, but not proven, to be flagellin.
Thus, it is clear that a need exists for a method of detecting Lyme borreliosis disease in mammals. The present invention provides such a method.